title: Labeling of primary human hepatocytes with micron-sized iron oxide particles in suspension culture suitable for large-scale preparation.
abstract: Labeling of hepatocytes with micron-sized iron oxide particles (MPIOs) enables cell detection using clinical magnetic resonance equipment. For clinical applications, large numbers of cells must be labeled in a simple and rapid manner and have to be applied in suspension. However, all existing protocols are based on adhesion culture labeling with subsequent resuspension, only suitable for small experimental settings. The aim of this study was to investigate the feasibility of preparing MPIO-labeled primary human hepatocytes in a temporary suspension culture. Human hepatocytes were isolated from 16 donors and labeled with MPIOs in suspension, using the Rotary Cell Culture System. Particle incorporation was investigated by light and electron microscopy. Cells were compared with adhesion culture-labeled and subsequently enzymatically resuspended cells. During a period of 5 days, hepatocyte-specific parameters of cell damage (aspartate aminotransferase and alanine aminotransferase) and metabolic activity (urea and albumin) were analyzed (n=7). Suspension cultures showed a higher outcome in cell recovery compared with the conventional labeling method. When incubated with 180 particles/viable cell for 4 h, the mean particle uptake was 28.8 particles/cell at a labeling efficiency of 95.1%. Labeling in suspension had no adverse effects on cell integrity or metabolic activity. We conclude that labeling of human hepatocytes in suspension is feasible and simple and may serve future large-scale processing of cells.
authors.affiliation: General, Visceral, and Transplantation Surgery, Experimental Surgery and Regenerative Medicine, Charité-Campus Virchow, Universitätsmedizin Berlin, Augustenburger Platz 1, Berlin, Germany. nora.kammer@charite.de
authors.forename: Nora N
authors.initials: NN
authors.lastname: Kammer
authors.forename: Nils
authors.initials: N
authors.lastname: Billecke
authors.forename: Mehmet H
authors.initials: MH
authors.lastname: Morgul
authors.forename: Michaela K
authors.initials: MK
authors.lastname: Adonopoulou
authors.forename: Martina
authors.initials: M
authors.lastname: Mogl
authors.forename: Mao D
authors.initials: MD
authors.lastname: Huang
authors.forename: Stefan
authors.initials: S
authors.lastname: Florek
authors.forename: Katharina R L
authors.initials: KR
authors.lastname: Schmitt
authors.forename: Nathanael
authors.initials: N
authors.lastname: Raschzok
authors.forename: Igor M
authors.initials: IM
authors.lastname: Sauer
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D018929:Cell Culture Techniques; D002470:Cell Survival; D002478:Cells, Cultured; D005290:Ferric Compounds / Q000032:analysis*; D022781:Hepatocytes / Q000166:cytology / Q000648:ultrastructure*; D006801:Humans; D008854:Microscopy, Electron; D008858:Microscopy, Phase-Contrast; D008875:Middle Aged; D010316:Particle Size; D013194:Staining and Labeling / Q000379:methods*; D055815:Young Adult
chemical_list: D005290:Ferric Compounds; C000499:ferric oxide